Phase Separations in Water-Salt Solutions of Polyelectrolyte Complexes Formed by RNA and Polycations: Comparison with DNA Complexes.
(2003) In Macromolecular Bioscience 3(8). p.404-411- Abstract
- Formation of insoluble polyelectrolyte complexes (PECs) between RNA and polycations was followed by measuring the residual RNA absorbance in the solution after separation of the precipitate. The polycations studied were poly(N,N-dimethyldiallylammonium) chloride (pendant type) and 2,5-ionene bromide (integral type) with quaternary amino groups in every monomer unit. The data obtained were compared with the results of analogous studies of DNA-containing PECs. This study is a part of a project aimed at the specific separation of plasmid DNA from RNA, a major problem in the preparative isolation of plasmid DNA. We thus deliberately chose a heterogenous RNA sample as it represents the RNA present in a real cell extract. In contrast to the... (More)
- Formation of insoluble polyelectrolyte complexes (PECs) between RNA and polycations was followed by measuring the residual RNA absorbance in the solution after separation of the precipitate. The polycations studied were poly(N,N-dimethyldiallylammonium) chloride (pendant type) and 2,5-ionene bromide (integral type) with quaternary amino groups in every monomer unit. The data obtained were compared with the results of analogous studies of DNA-containing PECs. This study is a part of a project aimed at the specific separation of plasmid DNA from RNA, a major problem in the preparative isolation of plasmid DNA. We thus deliberately chose a heterogenous RNA sample as it represents the RNA present in a real cell extract. In contrast to the exhaustive precipitation of DNA observed at certain values, a significant part of RNA was nonprecipitated at any = [+]/[-], that is, at any ratio of positively charged quaternary amino groups and negatively charged phosphate groups. The addition of sodium chloride increased the nonprecipitated fraction of RNA. DNA, on the other hand, was completely precipitated by both polycations at > 0.7. The less effective precipitation of RNA was probably due to the presence of a considerable fraction of short-chained molecules, incapable of forming a sufficient cooperative system of salt bonds with the polycation. This assumption was supported by a separate experiment, in which the precipitation behavior of RNA fractions of different molecular masses was investigated. The same tendency, while less pronounced, was also ascertained for PECs formed by polycations with DNA fractions of different molecular masses. The possibility of using the revealed differences between DNA and RNA behavior for effective precipitation procedure useful in bioseparation is discussed. The difference in the precipitation behavior of nucleic acids of different molecular masses means there is a possibility for developing an enzymatic assay for DNAase and RNAase activity. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/128638
- author
- Wahlund, Per-Olof LU ; Izumrudov, Vladimir LU ; Gustavsson, Per-Erik LU ; Larsson, Per-Olof LU and Galaev, Igor LU
- organization
- publishing date
- 2003
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Macromolecular Bioscience
- volume
- 3
- issue
- 8
- pages
- 404 - 411
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- wos:000184787300003
- scopus:0041968489
- ISSN
- 1616-5195
- DOI
- 10.1002/mabi.200350010
- language
- English
- LU publication?
- yes
- id
- 03395338-3979-46ab-bedc-f53a2f8feea0 (old id 128638)
- date added to LUP
- 2016-04-01 12:24:09
- date last changed
- 2022-01-27 03:16:23
@article{03395338-3979-46ab-bedc-f53a2f8feea0,
abstract = {{Formation of insoluble polyelectrolyte complexes (PECs) between RNA and polycations was followed by measuring the residual RNA absorbance in the solution after separation of the precipitate. The polycations studied were poly(N,N-dimethyldiallylammonium) chloride (pendant type) and 2,5-ionene bromide (integral type) with quaternary amino groups in every monomer unit. The data obtained were compared with the results of analogous studies of DNA-containing PECs. This study is a part of a project aimed at the specific separation of plasmid DNA from RNA, a major problem in the preparative isolation of plasmid DNA. We thus deliberately chose a heterogenous RNA sample as it represents the RNA present in a real cell extract. In contrast to the exhaustive precipitation of DNA observed at certain values, a significant part of RNA was nonprecipitated at any = [+]/[-], that is, at any ratio of positively charged quaternary amino groups and negatively charged phosphate groups. The addition of sodium chloride increased the nonprecipitated fraction of RNA. DNA, on the other hand, was completely precipitated by both polycations at > 0.7. The less effective precipitation of RNA was probably due to the presence of a considerable fraction of short-chained molecules, incapable of forming a sufficient cooperative system of salt bonds with the polycation. This assumption was supported by a separate experiment, in which the precipitation behavior of RNA fractions of different molecular masses was investigated. The same tendency, while less pronounced, was also ascertained for PECs formed by polycations with DNA fractions of different molecular masses. The possibility of using the revealed differences between DNA and RNA behavior for effective precipitation procedure useful in bioseparation is discussed. The difference in the precipitation behavior of nucleic acids of different molecular masses means there is a possibility for developing an enzymatic assay for DNAase and RNAase activity.}},
author = {{Wahlund, Per-Olof and Izumrudov, Vladimir and Gustavsson, Per-Erik and Larsson, Per-Olof and Galaev, Igor}},
issn = {{1616-5195}},
language = {{eng}},
number = {{8}},
pages = {{404--411}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Macromolecular Bioscience}},
title = {{Phase Separations in Water-Salt Solutions of Polyelectrolyte Complexes Formed by RNA and Polycations: Comparison with DNA Complexes.}},
url = {{http://dx.doi.org/10.1002/mabi.200350010}},
doi = {{10.1002/mabi.200350010}},
volume = {{3}},
year = {{2003}},
}